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dc.contributor.authorTarpey, I
dc.contributor.authorStacey, S N
dc.contributor.authorMcIndoe, A
dc.contributor.authorDavies, D H
dc.date.accessioned2010-04-06T09:46:11Z
dc.date.available2010-04-06T09:46:11Z
dc.date.issued1996-02
dc.identifier.citationPriming in vivo and quantification in vitro of class I MHC-restricted cytotoxic T cells to human papilloma virus type 11 early proteins (E6 and E7) using immunostimulating complexes (ISCOMs). 1996, 14 (3):230-6 Vaccineen
dc.identifier.issn0264-410X
dc.identifier.pmid8920705
dc.identifier.doi10.1016/0264-410X(95)00179-5
dc.identifier.urihttp://hdl.handle.net/10541/95623
dc.description.abstractImmunostimulating complexes (ISCOMs) efficiently deliver soluble antigen into both the cytosolic (endogenous) and endosomal (exogenous) pathways of antigen processing. Cytosolic delivery to antigen-presenting cells (APCs) may therefore be useful for the stimulation and assay of class I major histocompatibility complex (MHC)-restricted cytotoxic T lymphocytes (CTL) in vitro. In this study, mice were immunized with ISCOMs containing fusion proteins of the E6 or E7 early proteins of human papilloma virus type 11 (HPV 11) to elicit CTL. These CTL were then restimulated in vitro using APCs pulsed with the same ISCOMs, prior to cytotoxicity assay using syngeneic target cells infected with recombinant vaccinia viruses. In this way, antigen-specific, MHC-restricted lysis by CD8+ cells was detected. However, this was dependent on the use of low density splenocytes as APCs for restimulation in vitro. Limiting dilution analyses showed a direct correlation between the CTL responder frequency and the number of times the animals were immunized in vivo. We conclude that in lieu of infectious virus, the use of ISCOMs to mediate antigen delivery to APCs in vitro can be used to quantitate CTL activity. This may have applications in monitoring vaccine efficacy, particularly to viruses such as HPV, which cannot be presently obtained as infectious virus in sufficient quantity for CTL propagation and assay.
dc.language.isoenen
dc.subjectCultured Tumour Cellsen
dc.subject.meshAnimals
dc.subject.meshH-2 Antigens
dc.subject.meshISCOMs
dc.subject.meshMice
dc.subject.meshMice, Inbred C57BL
dc.subject.meshMice, Inbred DBA
dc.subject.meshOncogene Proteins, Viral
dc.subject.meshPapillomaviridae
dc.subject.meshSpleen
dc.subject.meshT-Lymphocytes, Cytotoxic
dc.subject.meshTumor Cells, Cultured
dc.subject.meshVaccines, Synthetic
dc.subject.meshViral Vaccines
dc.titlePriming in vivo and quantification in vitro of class I MHC-restricted cytotoxic T cells to human papilloma virus type 11 early proteins (E6 and E7) using immunostimulating complexes (ISCOMs).en
dc.typeArticleen
dc.contributor.departmentDivision of Life Sciences, King's College, London, UK.en
dc.identifier.journalVaccineen
html.description.abstractImmunostimulating complexes (ISCOMs) efficiently deliver soluble antigen into both the cytosolic (endogenous) and endosomal (exogenous) pathways of antigen processing. Cytosolic delivery to antigen-presenting cells (APCs) may therefore be useful for the stimulation and assay of class I major histocompatibility complex (MHC)-restricted cytotoxic T lymphocytes (CTL) in vitro. In this study, mice were immunized with ISCOMs containing fusion proteins of the E6 or E7 early proteins of human papilloma virus type 11 (HPV 11) to elicit CTL. These CTL were then restimulated in vitro using APCs pulsed with the same ISCOMs, prior to cytotoxicity assay using syngeneic target cells infected with recombinant vaccinia viruses. In this way, antigen-specific, MHC-restricted lysis by CD8+ cells was detected. However, this was dependent on the use of low density splenocytes as APCs for restimulation in vitro. Limiting dilution analyses showed a direct correlation between the CTL responder frequency and the number of times the animals were immunized in vivo. We conclude that in lieu of infectious virus, the use of ISCOMs to mediate antigen delivery to APCs in vitro can be used to quantitate CTL activity. This may have applications in monitoring vaccine efficacy, particularly to viruses such as HPV, which cannot be presently obtained as infectious virus in sufficient quantity for CTL propagation and assay.


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